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Investigations of Gluten Proteins from Functional and Historical Perspectives

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Investigations of Gluten Proteins from Functional and Historical Perspectives AN ABSTRACT OF THE THESIS OF Ying Zhang for the degree of Master of Science in Food Science and Technology presented on February 17, 2015. Title: Investigations of Gluten Proteins from Functional and Historical Perspectives Abstract approved: _____________________________________________________ Andrew S. Ross Wheat (Triticum aestivum) is one of the world’s major staple food crops, with flour produced from starchy endosperm being used for breads, cakes, noodles and various other wheat-based foods. The unique bread making properties of wheat are primarily attributed to its gluten-forming storage proteins: gliadins and glutenins. This study investigated the gluten proteins from functional and historical perspectives. The first study examined primarily the functional role of gluten proteins in the outcomes of the standard Falling Number (FN) test. The FN test is used in the grain trade to screen delivered wheat for the presence of pre-harvest sprouting by indirectly measuring α- amylase through it effects on the physical consistency of a cooked flour-water suspension. Grain protein content (GPC) has been implicated as a potential modifier of FN independent of α-amylase or sprout status. In the gluten functionality study, we proposed a protein unfolding and crosslinking model, and hypothesized that gluten proteins with higher molecular weight distributions (MWD) would heatset faster, tightly cover starch granules, restrict water entry, and slow their disintegration. In contrast to our hypothesis, our results showed that samples with lower MWD had faster heatset times than samples with higher MWD according to a controlled heating test. We also hypothesize that increased granularity of hard wheat flour reduces the surface area to volume ratio so the starch granules embedded in the particles need more time to hydrate or swell. However, our results indicated that natural variations in flour particle size from a standard grinding procedure that used a 0.8 mm screen had no impact on FN. The second study looked at potential changes in gluten proteins in a historical set of wheat varieties spanning more than 110 years of production. The wheats were in two sets: soft wheats where there has been no systematic selection for increased dough strength in breeding programs, and hard wheats where there has been a concerted effort to increase overall dough strength over the last century. The sample sets also covered the eras before and after the introduction of the semi-dwarf wheats to the USA. The reason for this investigation is related to the circumstance that wheat is the cause of celiac disease (CD) and is implcated in the disputed condition, non-celiac gluten sensitivity (NCGS). Recently, diagnoses of CD at least have increased and there are suggestions that changes in gluten proteins in the modern era are responsible. Since it is primarily gliadins that trigger CD, it was considered worth investigating whether or not there have been any changes in the composition of gliadins over the last century. Sixty-two soft and 61 hard U.S. high production wheat varieties from 1900 to the present (with one from 1800) were collected and analyzed by RP-HPLC. These varieties were investigated to begin to answer whether wheat breeding for higher dough strength, or the incorporation of dwarfing alleles after the 1960s, was associated with observable changes in gliadin composition. ANOVA showed that there was no significant difference between soft and hard wheats in the relative abundance of α/β-gliadins. However, there were significant differences between hard and soft wheats in the relative abundance of ω- and γ-gliadins. ANOVA also showed that there was no significant difference between tall and dwarf wheats in the relative abundance of any of the three gliadin fractions. The ANOVA results suggested that deliberate breeding for dough strength, as illustrated by the hard versus soft wheat contrast, had not systematically changed the relative abundance of α/β- gliadins across the last 110 years, but had altered the relative abundance of the other two fractions. ANOVA results indicated no change in proportions of the three gliadin fractions after deployment of the dwarfing alleles suggesting the tall to dwarf change was independent of gluten composition. Second order polynomial regression analyses showed that the relative abundance of α/β-gliadins increased until around 1960 then decreased. The changes were more noticeable in the hard wheats. The converse was observed for 훾- gliadins. This stepwise change questioned the association between CD increase and breeding for increased dough strength in hard wheats, since the relative abundance of α/β-gliadins did not keep going up, and α-gliadin is considered the major trigger force for CD initiation. In contrast, linear correlation analyses with each of 700, three second long fractions of the RP-HPLC chromatograms suggested that most changes were related to the soft wheat population. The discrepancy between the regression analyses of the three major fractions and the 700 small fractions may be related to the use of linear correlations in the latter when some relationships were clearly non-linear. Overall, our results did not fully support speculations that there have been profound changes in gluten composition related to the dwarfing alleles or selection for increased dough strength in hard wheats. ©Copyright by Ying Zhang February 17, 2015 All Rights Reserved Investigations of Gluten Proteins from Functional and Historical Perspectives by Ying Zhang A THESIS submitted to Oregon State University In partial fulfillment of the requirements for the degree of Master of Science Presented February 17, 2015 Commencement June 2015 Master of Science thesis of Ying Zhang presented on February 17, 2015 APPROVED: Major Professor, representing Food Science and Technology Head of the Department of Food Science and Technology Dean of the Graduate School I understand that my thesis will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my thesis to any reader upon request. Ying Zhang, Author ACKNOWLEDGEMENTS I would like to express my appreciation to all the people who contributed to the successful completion of this thesis. My advisor, Dr. Andrew Ross, deserves a very special acknowledgement for his patient guidance and assistance during this research. He gave me the opportunity to continue working on my interest area – instrumental analysis of biomolecules. I would like to thank Drs. Teepakorn Kongraksawech, Michael Penner, and Joseph McGuire for serving on my committee. I would also like to thank Colleen Roseborough for her help in materials supply and Dr. Jae Ohm for his time in data analysis. My sincere thanks to Caryn Ong, Dr. Teepakorn Kongraksawech, Mike Adams, Carlos Fajardo, Eva Kuhn, Joshua Evans, Omar Miranda-Garcia, Michayla Robertson and Jordan Smith in the Ross Lab. They provided tremendous help on this work. It was always a great time working with them and I am sure I will miss that. I would like to express my deepest gratitude towards my husband Kain Wu for his love, encouragement and unconditional support on every decision I made. Finally, I would like to thank my parents, whose infinite love and support made it possible for me to complete my study in OSU. CONTRIBUTION OF AUTHORS Dr. Andrew Ross and Dr. Robert Zemetra initiated the project. Dr. Andrew Ross guided the project and contributed to the manuscripts. Dr. Teepakorn Kongraksawech assisted with laboratory procedures. Dr. Jae Ohm performed the MATLAB interpolations of the RP-HPLC chromatogram data and assisted in their statistical interpretation. Colleen Roseborough conducted materials preparation for the historical study. TABLE OF CONTENTS Page Chapter 1: General Introduction..……………………………………………………1 1.1 Background……………………………………………………………….….1 1.2 Objectives and hypotheses…………….……………………………………..5 Chapter 2: Literature Review………………………………………………………...7 2.1 Wheat..…………………………………………………………….…………7 2.2 Wheat types and general uses.………….……………………………………8 2.3 Wheat kernel anatomy and composition…….………………………………10 2.4 Kernel physical texture………………………………………………………11 2.5 Wheat flour milling.…………………………………………………………13 2.6 Wheat proteins………………………………………………………………15 2.6.1 Gliadins………………………………………………………………….16 2.6.2 Glutenins...………………………………………………………………19 2.7 Wheat carbohydrates…….…………………………………………………..22 2.7.1 Starch...……………………………………………………………………22 2.7.2 Non-starch polysaccharides...……………………………………………28 2.8 Wheat lipids…….…………………………………………………...……….33 2.9 Wheat enzymes…….…………………………………………………...……34 2.9.1 Starch-degrading enzymes.………………………………………………35 2.9.2 Protein-degrading enzymes....……………………………………………36 2.9.3 Oxidoreductases………………………………………………………….37 2.10 The flour-water system during processing.…………………………………38 TABLE OF CONTENTS (Continued) Page 2.11 Gluten intolerance..…………………………………………………………42 References.………….……………………………………………………………44 Chapter 3: Investigations of Non-amylase Factors affecting Wheat Falling Number Results..…………………………………………………………45 Abstract………………..…………………………………………………………45 3.1 Introduction...………….………………………………..……………………45 3.2 Materials and methods..………………………………………………………49 3.2.1 Materials....…………….…………………………………………………49 3.2.2 Grain and flour physicochemical properties.…….……………………….51 3.2.3 Polymeric proteins analysis.…………….………………………………..52 3.2.4 Mixograph method...…….………………………………………….……56 3.2.5 Pasting properties..….……………………………………………………56 3.2.6 Particle size analysis...……………………………………………………58 3.2.7 Statistical
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